DOCSLIB.ORG

Identification of Uterine Ion Transporters for Mineralisation Precursors of the Avian Eggshell Vincent Jonchère, Aurélien Brionne, Joël Gautron and Yves Nys*

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Identification of Uterine Ion Transporters for Mineralisation Precursors of the Avian Eggshell Vincent Jonchère, Aurélien Brionne, Joël Gautron and Yves Nys* Jonchère et al. BMC Physiology 2012, 12:10 http://www.biomedcentral.com/1472-6793/12/10 RESEARCH ARTICLE Open Access Identification of uterine ion transporters for mineralisation precursors of the avian eggshell Vincent Jonchère, Aurélien Brionne, Joël Gautron and Yves Nys* Abstract Background: In Gallus gallus, eggshell formation takes place daily in the hen uterus and requires large amounts of 2+ - the ionic precursors for calcium carbonate (CaCO3). Both elements (Ca , HCO3) are supplied by the blood via trans-epithelial transport. Our aims were to identify genes coding for ion transporters that are upregulated in the uterine portion of the oviduct during eggshell calcification, compared to other tissues and other physiological states, and incorporate these proteins into a general model for mineral transfer across the tubular gland cells during eggshell formation. Results: A total of 37 candidate ion transport genes were selected from our database of overexpressed uterine genes associated with eggshell calcification, and by analogy with mammalian transporters. Their uterine expression was compared by qRTPCR in the presence and absence of eggshell formation, and with relative expression levels in 2+ - 2+ - magnum (low Ca /HCO3 movement) and duodenum (high rates of Ca /HCO3 trans-epithelial transfer). We identified overexpression of eleven genes related to calcium movement: the TRPV6 Ca2+ channel (basolateral uptake of Ca2+), 28 kDa calbindin (intracellular Ca2+ buffering), the endoplasmic reticulum type 2 and 3 Ca2+ pumps (ER uptake), and the inositol trisphosphate receptors type 1, 2 and 3 (ER release). Ca2+ movement across the apical membrane likely involves membrane Ca2+ pumps and Ca2+/Na+ exchangers. Our data suggests that Na+ transport involved the SCNN1 channel and the Na+/Ca2+ exchangers SLC8A1, 3 for cell uptake, the Na+/K+ ATPase for cell output. K+ uptake resulted from the Na+/K+ ATPase, and its output from the K+ channels (KCNJ2, 15, 16 and KCNMA1). - - We propose that the HCO3 is mainly produced from CO2 by the carbonic anhydrase 2 (CA2) and that HCO3 is - - - 2+ + secreted through the HCO3/Cl exchanger SLC26A9. HCO3 synthesis and precipitation with Ca produce two H . Protons are absorbed via the membrane’sCa2+ pumps ATP2B1, 2 in the apical membrane and the vacuolar - - - (H+)-atpases at the basolateral level. Our model incorporate Cl ions which are absorbed by the HCO3/Cl exchanger SLC26A9 and by Cl- channels (CLCN2, CFTR) and might be extruded by Cl-/H+ exchanger (CLCN5), but also by Na+ K+ 2Cl- and K+ Cl- cotransporters. Conclusions: Our Gallus gallus uterine model proposes a large list of ion transfer proteins supplying Ca2+ and - HCO3 and maintaining cellular ionic homeostasis. This avian model should contribute towards understanding the mechanisms and regulation for ionic precursors of CaCO3, and provide insight in other species where epithelia transport large amount of calcium or bicarbonate. Keywords: Ion, Mineral, Calcium, Transporter, Uterus, Eggshell, Chicken * Correspondence: [email protected] INRA, UR83 Recherches Avicoles, F-37380, Nouzilly, France © 2012 Jonchère et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Jonchère et al. BMC Physiology 2012, 12:10 Page 2 of 17 http://www.biomedcentral.com/1472-6793/12/10 Background uterine calbindin levels and Ca2+ flux [11,13,14]. This Biomineralisation is a process by which living organisms protein could also take part in maintaining low intracel- develop mineral structures to perform a variety of roles lular Ca2+ to avoid cell death as observed in other spe- related to support, defence and feeding. Amongst these, cies and tissues [15]. Ca2+ secretion from epithelial cells a large number of animals (birds, molluscs, foraminifera, to the uterine fluid is active involving a Ca2+ ATPase, corals, sea urchins) mineralises by co-precipitation of the activity of which varies with the stage of eggshell cal- 2+ 2- calcium (Ca ) and carbonates (CO3 ) to form a protect- cification [4,7]. A recent study [16] identified and loca- ive shell or a skeleton. The prerequisite for shell min- lized the plasma membrane Ca2+ ATPase isoform 4 eralisation is the supply of large amounts of Ca2+ and (PMCA4) in the apical membrane of epithelial cells of 2- + CO3 in a limited extracellular milieu by trans-cellular king quail. The disruption of sodium (Na ) re- transport, requiring the presence of ion channels, ion absorption by specific inhibitors in perfused uterus or pumps and ion exchangers. In Gallus gallus, eggshell in vitro reduced Ca2+ secretion by 50% [9,17], revealing formation takes place daily in the hen uterus and is one a strong relationship between Na+ and Ca2+ transfers of the most rapid mineralisation processes [1]. It and therefore the putative presence of Na+/Ca2+ exchan- + + requires large amount of calcium carbonate (CaCO3)as gers in uterine cells. The Na /K ATPase responsible for the hen exports the equivalent of her body weight as Na+ re-absorption in the plasma membrane is charac- eggshell in one year of egg production (>1.5 kg). Both terised and is upregulated during the period of shell 2+ - elements (Ca and HCO3) are not stored in the uterus calcification [18]. but are continuously supplied during eggshell formation The second essential component of eggshell mineral- by the blood plasma via trans-epithelial transport taking isation is carbonate. Blood carbon dioxide (CO2) is pro- place across the uterine glandular cells [2-4]. Early stud- vided in cells by passive diffusion through the plasma ies determined the ion concentrations of the uterine membrane [2,19]. In the uterine tubular gland cells, a fluid, which bathes the eggshell and changes during the family of key enzymes, the carbonic anhydrases (CA) [6] - sequential stages of calcification (Table 1) [5], identified catalyses the hydration of CO2 to HCO3 as confirmed by - several proteins involved in ion transport [3,6,7], and inhibition of HCO3production and secretion by acetazo- recorded changes in ion fluxes across the uterine epithe- lamide, a CA inhibitor [9]. Chloride (Cl-) is absorbed by lium in response to ion transporter inhibitors [8-10]. the uterus and any perturbation of Na+ flux by ouabain These classic approaches led to a hypothesis concerning [9] reverses both the Na+ and Cl- fluxes, but reduces also - the mechanisms of ion transfer through the uterine glan- HCO3 secretion suggesting that its transfer is dependent 2+ - - - dular cells (Figure 1; [1]). In hens, the Ca blood on Cl via a Cl /HCO3 exchanger which has not been -4 - (1.2 mM) and epithelial cell concentrations (10 mM), identified. Finally, the production of HCO3 in tubular 2+ 2+ 2- suggest that Ca entry in cell is passive via a Ca chan- gland cells and of CO3 in the uterine fluid generates nel, which remains unidentified. The intracellular Ca2+ high levels of protons (H+) ions. The concomitant de- transport through the cell involves 28 kDa calbindin crease in uterine and plasma pH during calcification [3,11,12]. The 28 kDa calbindin expression is greatly reflects the reabsorption of H+ [5]. upregulated during eggshell formation and falls after Only a few genes and related proteins involved in uter- suppression of calcification (by premature egg expul- ine ion transfer have been identified to date. Our object- sion), suggesting a very close relationship between ive therefore was to use the recent information issuing from the chicken genome sequencing [20] and subse- Table 1 PH and ion concentrations in blood plasma, quent enrichment in the chicken gene/protein databases uterine fluid and epithelial cells during eggshell to identify uterine ion transport proteins. Use of a recent mineralisation[5] transcriptomic study revealing uterine genes related to Blood plasma Epithelial cells Uterine fluid eggshell calcification [21] and of the analogies with 8 h PO 18 h PO transporters previously described in mammalian tissues Ions [mM] [mM] [mM] [mM] transferring large quantities of ions (intestine, kidney, Ca2+ 1.2 <0.0002 6 10 pancreas) allows the identification of putative genes en- Na+ 140 12 144 80 coding proteins involved in uterine trans-epithelial ion K+ 4 139 12 60 transports. Confirmation of their presence in birds and - evaluation of their involvement have been analysed by HCO3 23 12 60 110 comparing gene expression in the uterus compared to pH 7.4 7.0-7.4 7.6 7.1 the magnum (the oviduct segment responsible for the Cl- 130 4 71 45 synthesis and secretion of egg white proteins) and the The eggshell precursors are secreted in the uterine fluid where the eggshell 2+ mineralization daily takes place from 10 hours to 22 hours post ovulation (yolk duodenum (Ca uptake and neutralization of stomach 2+ - entry in the oviduct). PO: post ovulation. acid), where both Ca and HCO3 trans-epithelial Jonchère et al. BMC Physiology 2012, 12:10 Page 3 of 17 http://www.biomedcentral.com/1472-6793/12/10 Figure 1 Classic hypothesis concerning ion transfers in the hen uterus during eggshell calcification [1,5,8-10]. Ca2+ entry in cell is passive via a Ca2+ channel, 28 kDa calbindin contributes to intracellular transfer and maintenance of a low Ca2+ level. Ca2+ secretion involving a Ca2+ 2+ + ATPase and a Ca /Na exchanger. Carbonic anhydrase has a key role in providing carbonate from plasma CO2. transfers are respectively low and high. The magnum to the magnum (egg white protein secretion) [21] and and the uterus secrete a large amount of water, Na+ and analogies with transporters previously described in Cl- during the phase of hydration of egg albumen which mammalian tissues at the intestinal and kidney level takes place before the active phase of eggshell formation [24,26].
Load more

Recommended publications
  • Molecular Profile of Tumor-Specific CD8+ T Cell Hypofunction in a Transplantable Murine Cancer Model
    Downloaded from http://www.jimmunol.org/ by guest on September 25, 2021 T + is online at: average * The Journal of Immunology , 34 of which you can access for free at: 2016; 197:1477-1488; Prepublished online 1 July from submission to initial decision 4 weeks from acceptance to publication 2016; doi: 10.4049/jimmunol.1600589 http://www.jimmunol.org/content/197/4/1477 Molecular Profile of Tumor-Specific CD8 Cell Hypofunction in a Transplantable Murine Cancer Model Katherine A. Waugh, Sonia M. Leach, Brandon L. Moore, Tullia C. Bruno, Jonathan D. Buhrman and Jill E. Slansky J Immunol cites 95 articles Submit online. Every submission reviewed by practicing scientists ? is published twice each month by Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts http://jimmunol.org/subscription Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html http://www.jimmunol.org/content/suppl/2016/07/01/jimmunol.160058 9.DCSupplemental This article http://www.jimmunol.org/content/197/4/1477.full#ref-list-1 Information about subscribing to The JI No Triage! Fast Publication! Rapid Reviews! 30 days* Why • • • Material References Permissions Email Alerts Subscription Supplementary The Journal of Immunology The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2016 by The American Association of Immunologists, Inc. All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. This information is current as of September 25, 2021. The Journal of Immunology Molecular Profile of Tumor-Specific CD8+ T Cell Hypofunction in a Transplantable Murine Cancer Model Katherine A.
    [Show full text]
  • Plasma Membrane Ca2+–Atpase in Rat and Human Odontoblasts Mediates Dentin Mineralization
    biomolecules Article Plasma Membrane Ca2+–ATPase in Rat and Human Odontoblasts Mediates Dentin Mineralization Maki Kimura 1,†, Hiroyuki Mochizuki 1,†, Ryouichi Satou 2, Miyu Iwasaki 2, Eitoyo Kokubu 3, Kyosuke Kono 1, Sachie Nomura 1, Takeshi Sakurai 1, Hidetaka Kuroda 1,4,† and Yoshiyuki Shibukawa 1,*,† 1 Department of Physiology, Tokyo Dental College, 2-9-18, Kanda-Misaki-cho, Chiyoda-ku, Tokyo 101-0061, Japan; [email protected] (M.K.); [email protected] (H.M.); [email protected] (K.K.); [email protected] (S.N.); [email protected] (T.S.); [email protected] (H.K.) 2 Department of Epidemiology and Public Health, Tokyo Dental College, Chiyodaku, Tokyo 101-0061, Japan; [email protected] (R.S.); [email protected] (M.I.) 3 Department of Microbiology, Tokyo Dental College, Chiyodaku, Tokyo 101-0061, Japan; [email protected] 4 Department of Dental Anesthesiology, Kanagawa Dental University, 1-23, Ogawacho, Kanagawa, Yokosuka-shi 238-8570, Japan * Correspondence: [email protected] † These authors contributed equally to this study. Abstract: Intracellular Ca2+ signaling engendered by Ca2+ influx and mobilization in odontoblasts is critical for dentinogenesis induced by multiple stimuli at the dentin surface. Increased Ca2+ is exported by the Na+–Ca2+ exchanger (NCX) and plasma membrane Ca2+–ATPase (PMCA) to Citation: Kimura, M.; Mochizuki, H.; maintain Ca2+ homeostasis. We previously demonstrated a functional coupling between Ca2+ Satou, R.; Iwasaki, M.; Kokubu, E.; extrusion by NCX and its influx through transient receptor potential channels in odontoblasts. Kono, K.; Nomura, S.; Sakurai, T.; Although the presence of PMCA in odontoblasts has been previously described, steady-state levels of Kuroda, H.; Shibukawa, Y.
    [Show full text]
  • To Study Mutant P53 Gain of Function, Various Tumor-Derived P53 Mutants
    Differential effects of mutant TAp63γ on transactivation of p53 and/or p63 responsive genes and their effects on global gene expression. A thesis submitted in partial fulfillment of the requirements for the degree of Master of Science By Shama K Khokhar M.Sc., Bilaspur University, 2004 B.Sc., Bhopal University, 2002 2007 1 COPYRIGHT SHAMA K KHOKHAR 2007 2 WRIGHT STATE UNIVERSITY SCHOOL OF GRADUATE STUDIES Date of Defense: 12-03-07 I HEREBY RECOMMEND THAT THE THESIS PREPARED UNDER MY SUPERVISION BY SHAMA KHAN KHOKHAR ENTITLED Differential effects of mutant TAp63γ on transactivation of p53 and/or p63 responsive genes and their effects on global gene expression BE ACCEPTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF Master of Science Madhavi P. Kadakia, Ph.D. Thesis Director Daniel Organisciak , Ph.D. Department Chair Committee on Final Examination Madhavi P. Kadakia, Ph.D. Steven J. Berberich, Ph.D. Michael Leffak, Ph.D. Joseph F. Thomas, Jr., Ph.D. Dean, School of Graduate Studies 3 Abstract Khokhar, Shama K. M.S., Department of Biochemistry and Molecular Biology, Wright State University, 2007 Differential effect of TAp63γ mutants on transactivation of p53 and/or p63 responsive genes and their effects on global gene expression. p63, a member of the p53 gene family, known to play a role in development, has more recently also been implicated in cancer progression. Mice lacking p63 exhibit severe developmental defects such as limb truncations, abnormal skin, and absence of hair follicles, teeth, and mammary glands. Germline missense mutations of p63 have been shown to be responsible for several human developmental syndromes including SHFM, EEC and ADULT syndromes and are associated with anomalies in the development of organs of epithelial origin.
    [Show full text]
  • Optimisation and Validation of a High Throughput Screening Compatible
    J Pharm Pharmaceut Sci (www.cspsCanada.org) 16(2) 217 - 230, 2013 Optimisation and Validation of a High Throughput Screening Compatible Assay to Identify Inhibitors of the Plasma Membrane Calcium ATPase Pump - a Novel Therapeutic Target for Contraception and Malaria Tamer M A Mohamed1,2, Simon A Zakeri1, Florence Baudoin1, Markus Wolf3, Delvac Oceandy1, Elizabeth J Cartwright1, Sheraz Gul3, Ludwig Neyses1 1 Institute of Cardiovascular Sciences, Manchester Academic Health Sciences Centre, University of Manchester, Manchester M13 9PT, UK. 2 Dept. of Biochemistry, Faculty of Pharmacy, Zagazig University, Zagazig, Egypt. 3 European ScreeningPort GmbH, 22525 Hamburg, Germany. Received, March 13, 2013; Revised, April 25, 2013; Accepted, May 8th, 2013; Published, May 10th, 2013. ABSTRACT – Purpose. ATPases, which constitute a major category of ion transporters in the human body, have a variety of significant biological and pathological roles. However, the lack of high throughput assays for ATPases has significantly limited drug discovery in this area. We have recently found that the genetic deletion of the ATP dependent calcium pump PMCA4 (plasma membrane calcium/calmodulin dependent ATPase, isoform 4) results in infertility in male mice due to a selective defect in sperm motility. In addition, recent discoveries in humans have indicated that a single nucleotide polymorphism (SNP) in the PMCA4 gene determines the susceptibility towards malaria plasmodium infection. Therefore, there is an urgent need to develop specific PMCA4 inhibitors. In the current study, we aim to optimise and validate a high throughput screening compatible assay using recombinantly expressed PMCA4 and the HTRF® Transcreener® ADP (TR-FRET) assay to screen a drug library. Methods and Results.
    [Show full text]
  • Transcriptomic and Proteomic Profiling Provides Insight Into
    BASIC RESEARCH www.jasn.org Transcriptomic and Proteomic Profiling Provides Insight into Mesangial Cell Function in IgA Nephropathy † † ‡ Peidi Liu,* Emelie Lassén,* Viji Nair, Celine C. Berthier, Miyuki Suguro, Carina Sihlbom,§ † | † Matthias Kretzler, Christer Betsholtz, ¶ Börje Haraldsson,* Wenjun Ju, Kerstin Ebefors,* and Jenny Nyström* *Department of Physiology, Institute of Neuroscience and Physiology, §Proteomics Core Facility at University of Gothenburg, University of Gothenburg, Gothenburg, Sweden; †Division of Nephrology, Department of Internal Medicine and Department of Computational Medicine and Bioinformatics, University of Michigan, Ann Arbor, Michigan; ‡Division of Molecular Medicine, Aichi Cancer Center Research Institute, Nagoya, Japan; |Department of Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden; and ¶Integrated Cardio Metabolic Centre, Karolinska Institutet Novum, Huddinge, Sweden ABSTRACT IgA nephropathy (IgAN), the most common GN worldwide, is characterized by circulating galactose-deficient IgA (gd-IgA) that forms immune complexes. The immune complexes are deposited in the glomerular mesangium, leading to inflammation and loss of renal function, but the complete pathophysiology of the disease is not understood. Using an integrated global transcriptomic and proteomic profiling approach, we investigated the role of the mesangium in the onset and progression of IgAN. Global gene expression was investigated by microarray analysis of the glomerular compartment of renal biopsy specimens from patients with IgAN (n=19) and controls (n=22). Using curated glomerular cell type–specific genes from the published literature, we found differential expression of a much higher percentage of mesangial cell–positive standard genes than podocyte-positive standard genes in IgAN. Principal coordinate analysis of expression data revealed clear separation of patient and control samples on the basis of mesangial but not podocyte cell–positive standard genes.
    [Show full text]
  • 1.7 the Plasma Membrane Calcium Atpase (PMCA)
    University of Manchester The role of PMCA1 in post- myocardial infarction remodelling A thesis submitted to the University of Manchester for the degree of Doctor of Philosophy in the Faculty of Biology, Medicine and Health 2017 Vera Stankovikj School of Medical Sciences Division of Cardiovascular Sciences Table of contents List of figures ....................................................................................................................... 10 List of tables ........................................................................................................................ 14 List of abbreviations ............................................................................................................. 16 Declaration .......................................................................................................................... 20 Copyright statement ............................................................................................................ 21 Abstract ............................................................................................................................... 22 Acknowledgements ............................................................................................................. 24 1. Introduction ................................................................................................................. 26 1.1 Heart failure ........................................................................................................... 26 1.2 Coronary artery
    [Show full text]
  • Serum from Patients with Hypertension Promotes Endothelial Dysfunction
    MOLECULAR MEDICINE REPORTS 23: 319, 2021 Serum from patients with hypertension promotes endothelial dysfunction to induce trophoblast invasion through the miR‑27b‑3p/ATPase plasma membrane Ca2+ transporting 1 axis LIBO ZHU and ZHUQING LIU Department of Obstetrics, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, P.R. China Received November 14, 2019; Accepted August 8, 2020 DOI: 10.3892/mmr.2021.11958 Abstract. Pregnancy‑induced hypertension is often miR‑27b‑3p mimics reduced the expression level of ATP2B1, accompanied by preeclampsia. The present study investigated and miR‑27b‑3p inhibitor reversed the effect of hypertensive whether microRNA (miR)‑27b‑3p affected the occurrence serum on ATP2B1 expression. Furthermore, patients with of preeclampsia by regulating the function of endothelial hypertension showed increased endothelial dysfunction, cells. Expressions levels of miR‑27b‑3p and ATPase plasma reduced trophoblastic invasion and the expressions of VEGF, membrane Ca2+ transporting 1 (ATP2B1) were determined MMP‑2 and MMP‑9, and miR‑27b‑3p mimics and silencing using reverse‑transcription quantitative PCR. miR‑27b‑3p of ATP2B1 produced similar results to HUVECs. The targeting ATP2B1 was predicted using bioinformatics and miR‑27b‑3p inhibitor reversed the effect of hypertensive further confirmed by dual‑luciferase reporter assays. Cell serum, and silencing of ATP2B1 inhibited the improvement Counting Kit‑8, Transwell and Matrigel tube formation of miR‑27b‑3p inhibitor to HUVECs and HTR‑8/SVneo cells assays were performed to detect the effects of miR‑27b‑3p on in proliferation, migration and tube formation. The current proliferation, migration and tube formation of human umbilical findings suggested that miR‑27b‑3p promoted proliferation, vein endothelial cells (HUVECs), respectively.
    [Show full text]
  • ELMO1 Signaling Is a Promoter of Osteoclast Function and Bone Loss ✉ Sanja Arandjelovic 1 , Justin S
    ARTICLE https://doi.org/10.1038/s41467-021-25239-6 OPEN ELMO1 signaling is a promoter of osteoclast function and bone loss ✉ Sanja Arandjelovic 1 , Justin S. A. Perry 1, Ming Zhou 2, Adam Ceroi3, Igor Smirnov4, Scott F. Walk1, Laura S. Shankman1, Isabelle Cambré3, Suna Onengut-Gumuscu 5, Dirk Elewaut 3, Thomas P. Conrads 2 & ✉ Kodi S. Ravichandran 1,3 Osteoporosis affects millions worldwide and is often caused by osteoclast induced bone loss. 1234567890():,; Here, we identify the cytoplasmic protein ELMO1 as an important ‘signaling node’ in osteoclasts. We note that ELMO1 SNPs associate with bone abnormalities in humans, and that ELMO1 deletion in mice reduces bone loss in four in vivo models: osteoprotegerin deficiency, ovariectomy, and two types of inflammatory arthritis. Our transcriptomic analyses coupled with CRISPR/Cas9 genetic deletion identify Elmo1 associated regulators of osteoclast function, including cathepsin G and myeloperoxidase. Further, we define the ‘ELMO1 inter- actome’ in osteoclasts via proteomics and reveal proteins required for bone degradation. ELMO1 also contributes to osteoclast sealing zone on bone-like surfaces and distribution of osteoclast-specific proteases. Finally, a 3D structure-based ELMO1 inhibitory peptide reduces bone resorption in wild type osteoclasts. Collectively, we identify ELMO1 as a signaling hub that regulates osteoclast function and bone loss, with relevance to osteoporosis and arthritis. 1 Center for Cell Clearance, Department of Microbiology, Immunology, and Cancer Biology and Carter Immunology Center, University of Virginia, Charlottesville, VA, USA. 2 Inova Schar Cancer Institute, Inova Center for Personalized Health, Fairfax, VA, USA. 3 Inflammation Research Centre, VIB, and the Department of Biomedical Molecular Biology, Ghent Univeristy, Ghent, Belgium.
    [Show full text]
  • The Effects of Anti-Hypertensive Drugs and the Mechanism of Hypertension in Vascular Smooth Muscle Cell-Specific ATP2B1 Knockout Mice
    Hypertension Research (2018) 41, 80–87 Official journal of the Japanese Society of Hypertension OPEN www.nature.com/hr ORIGINAL ARTICLE The effects of anti-hypertensive drugs and the mechanism of hypertension in vascular smooth muscle cell-specific ATP2B1 knockout mice Yuki Okuyama1, Nobuhito Hirawa2, Megumi Fujita1, Akira Fujiwara2, Yosuke Ehara1, Keisuke Yatsu1, Koichiro Sumida1, Minako Kagimoto1, Mari Katsumata2, Yusuke Kobayashi1, Sanae Saka2, Satoshi Umemura1 and Kouichi Tamura1 ATP2B1 is a gene associated with hypertension. We reported previously that mice lacking ATP2B1 in vascular smooth muscle cells (VSMC ATP2B1 KO mice) exhibited high blood pressure and increased intracellular calcium concentration. The present study was designed to investigate whether lack of the ATP2B1 gene causes a higher response to calcium channel blockers (CCBs) than to other types of anti-hypertensive drugs. Both VSMC ATP2B1 KO and control mice were administered anti- hypertensive drugs while monitoring blood pressure shifts. We also examined the association of nitric oxide synthase (NOS) activity in those mice to investigate whether another mechanism of hypertension existed. VSMC ATP2B1 KO mice exhibited significantly greater anti-hypertensive effects with a single injection of nicardipine, but the effects of an angiotensin II receptor blocker (ARB), an α-blocker and amlodipine on blood pressure were all similar to control mice. However, long-term treatment with amlodipine, but not an ARB, significantly decreased the blood pressure of KO mice compared with control mice. Both mRNA and protein expression levels of the L-type calcium channel were significantly upregulated in KO VSMCs. There were no alterations in neural NOS protein expression of VSMCs or in urinary NO production between the two groups.
    [Show full text]
  • Lncrna SNHG8 Is Identified As a Key Regulator of Acute Myocardial
    Zhuo et al. Lipids in Health and Disease (2019) 18:201 https://doi.org/10.1186/s12944-019-1142-0 RESEARCH Open Access LncRNA SNHG8 is identified as a key regulator of acute myocardial infarction by RNA-seq analysis Liu-An Zhuo, Yi-Tao Wen, Yong Wang, Zhi-Fang Liang, Gang Wu, Mei-Dan Nong and Liu Miao* Abstract Background: Long noncoding RNAs (lncRNAs) are involved in numerous physiological functions. However, their mechanisms in acute myocardial infarction (AMI) are not well understood. Methods: We performed an RNA-seq analysis to explore the molecular mechanism of AMI by constructing a lncRNA-miRNA-mRNA axis based on the ceRNA hypothesis. The target microRNA data were used to design a global AMI triple network. Thereafter, a functional enrichment analysis and clustering topological analyses were conducted by using the triple network. The expression of lncRNA SNHG8, SOCS3 and ICAM1 was measured by qRT-PCR. The prognostic values of lncRNA SNHG8, SOCS3 and ICAM1 were evaluated using a receiver operating characteristic (ROC) curve. Results: An AMI lncRNA-miRNA-mRNA network was constructed that included two mRNAs, one miRNA and one lncRNA. After RT-PCR validation of lncRNA SNHG8, SOCS3 and ICAM1 between the AMI and normal samples, only lncRNA SNHG8 had significant diagnostic value for further analysis. The ROC curve showed that SNHG8 presented an AUC of 0.850, while the AUC of SOCS3 was 0.633 and that of ICAM1 was 0.594. After a pairwise comparison, we found that SNHG8 was statistically significant (P SNHG8-ICAM1 = 0.002; P SNHG8-SOCS3 = 0.031).
    [Show full text]
  • Plasma Membrane Calcium Atpase Regulates Stoichiometry of CD4+ T-Cell Compartments
    ORIGINAL RESEARCH published: 21 May 2021 doi: 10.3389/fimmu.2021.687242 Plasma Membrane Calcium ATPase Regulates Stoichiometry of CD4+ T-Cell Compartments Maylin Merino-Wong, Barbara A. Niemeyer and Dalia Alansary* Molecular Biophysics, Saarland University, Homburg, Germany Immune responses involve mobilization of T cells within naïve and memory compartments. Tightly regulated Ca2+ levels are essential for balanced immune outcomes. How Ca2+ contributes to regulating compartment stoichiometry is unknown. Here, we show that plasma membrane Ca2+ ATPase 4 (PMCA4) is differentially expressed in human CD4+ T compartments yielding distinct store operated Ca2+ entry (SOCE) profiles. Modulation of PMCA4 yielded a more prominent increase of SOCE in memory than in naïve CD4+ T cell. Interestingly, downregulation of PMCA4 reduced the effector compartment fraction and Edited by: led to accumulation of cells in the naïve compartment. In silico analysis and chromatin Andrew D. Wells, immunoprecipitation point towards Ying Yang 1 (YY1) as a transcription factor regulating Children’s Hospital of Philadelphia, PMCA4 expression. Analyses of PMCA and YY1 expression patterns following activation United States and of PMCA promoter activity following downregulation of YY1 highlight repressive role of Reviewed by: fi 2+ Tomasz Boczek, YY1 on PMCA expression. Our ndings show that PMCA4 adapts Ca levels to cellular Boston Children’s Hospital and requirements during effector and quiescent phases and thereby represent a potential Harvard Medical School, United States target to intervene with the outcome of the immune response. Kai Yang, Indiana University School of Keywords: Plasma Membrane Calcium ATPase, Yin Yang 1, CD4+ compartments, calcium signaling, stoichiometry Medicine-Lafayette, United States *Correspondence: Dalia Alansary [email protected] INTRODUCTION 2+ Specialty section: The spatiotemporal characteristics of Ca signals in T cells tightly control the outcome of an This article was submitted to immune response as well as the fate of the immune cells involved.
    [Show full text]
  • The Effect of Ablation and Acute Inhibition Of
    The effect of ablation and acute inhibition of plasma membrane calcium ATPase 4 (PMCA4) with a novel inhibitor on isolated mouse mesenteric resistance arterial contractility A thesis submitted to The University of Manchester for the degree of Doctor of Philosophy (PhD) in the Faculty of Medical and Human Sciences Year of Submission: 2012 Sophronia Amabel Lewis School of Medicine Table of Contents Abstract ................................................................................................................ 13 Chapter 1 Introduction ........................................................................................ 27 1.1 Impact of cardiovascular diseases (CVD) in the UK .................................. 27 1.2 The importance of the resistance arteries in the cardiovascular system ...... 29 1.3 Regulation of arterial diameter .................................................................. 33 1.3.1 Mechanisms underlying modulation of VSM contractility; the role of calcium ……. ........................................................................................................ 33 1.3.2 Role of the endothelium in the regulation of arterial contractility ............. 37 1.3.3 Sources of NO and NO-mediated signalling in the vasculature ................. 39 1.3.4 Structural remodelling of resistance arteries ............................................. 41 1.4 Ca2+ mobilisation in VSM cells ................................................................. 43 1.4.1 Ca2+ mobilisation: Ca2+ entry across the plasma membrane
    [Show full text]